SENSITIVE NORTHERN BLOT HYBRIDIZATION USING DIGOXIGENIN RNA PROBES FOR THE MESSENGER-RNA DETECTION OF 2 GLUCOSE-TRANSPORTER ISOFORMS

Diseases involving glucose metabolism disorders are more and more prev alent. Therefore the question of glucose transporter gene expression i s being addressed in experimental and clinical studies. Radioactive pr obes are generally used to assess glucose transporter mRNA levels, but these probes are short-lived, costly and harmful to the environment. Alternative methods that do not present these disadvantages, for examp le digoxigenin (DIG) labelled probes, might prove to be very interesti ng for the study of glucose transporter mRNA. The aim of the present w ork was to compare DIG-labelled cRNA probes to P-32-labelled cRNA prob es in order to see whether or not the non-radioactive method can be us ed to assess glucose transporter gene expression. This work shows that DIG-labelled glucose transporter (GLUT1 and GLUT4) cRNAs are suitable probes for the assessment of these gene expressions. We have found th at the DIG system offers a much higher sensitivity than the P-32 syste m for both GLUT1 and GLUT4 mRNA detection. This represents a decisive advantage in human studies where tissue quantity is a limiting factor. In addition, stability, safety, time saving and cost reduction are ot her considerations that make DIG-labelled GLUT1 and GLUT4 cRNAs very a ttractive.